{"product_id":"e-coli-dna-topoisomerase-ii-gyrase-assay-kit-bht16300067","title":"E. coli DNA Topoisomerase II (Gyrase) Assay Kit","description":"\u003cstyle\u003e.bhp-desc{font-size:16px;color:#1a1a1a;line-height:1.7}.bhp-desc h2{font-size:18px;font-weight:700;color:#003366;margin:24px 0 10px;padding-bottom:6px;border-bottom:2px solid #003366}.bhp-desc p{margin:0 0 12px}.bhp-desc .bhp-spec-table{width:100%;border-collapse:collapse;margin:12px 0 18px}.bhp-desc .bhp-spec-table th{text-align:left;padding:8px 12px;background:#e8f0fb;color:#003366;font-weight:600;vertical-align:top;border:1px solid #ccd9f0}.bhp-desc .bhp-spec-table td{padding:8px 12px;vertical-align:top;border:1px solid #ccd9f0}.bhp-desc .bhp-warning-box{background:#fff8e6;border-left:4px solid #e6a817;padding:10px 14px;margin:10px 0 14px;border-radius:0 4px 4px 0}.bhp-desc .bhp-cat-pill{display:inline-block;background:#003366;color:#fff;border-radius:20px;padding:3px 12px;font-size:13px;margin-bottom:10px}.bhp-desc ol,.bhp-desc ul{margin:0 0 12px 20px}\u003c\/style\u003e\u003cdiv class=\"bhp-desc\"\u003e\n\u003cspan class=\"bhp-cat-pill\"\u003eDrug Target Assay Kit\u003c\/span\u003e\u003cp\u003eBacterial DNA topoisomerase II (gyrase) converts relaxed circular DNA into supercoiled DNA (the DNA supercoiling reaction). The change in fluorescence is used to measure gyrase supercoiling and to screen gyrase inhibitors. Inhibition of E. coli gyrase by novobiocin, ciprofloxacin, enrofloxacin and levofloxacin is shown in the protocol.\u003c\/p\u003e\n\u003ch2\u003eKit vs Kit Plus\u003c\/h2\u003e\n\u003cp\u003eDSA100K is the Kit without enzyme; DSA100KE (Kit Plus) adds 50 µl of 100 x E. coli gyrase. The H19 dye stock also differs: 8 µl of 4500 x dye in the Kit versus 20 µl of 1500 x dye in the Kit Plus; other components are the same, for 100 assays each.\u003c\/p\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eKit (enzyme not included), 100 assays (DSA100K)\u003c\/th\u003e\n\u003ctd\u003e100 assays\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eKit Plus (enzyme included), 100 assays (DSA100KE)\u003c\/th\u003e\n\u003ctd\u003e100 assays\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003ch2\u003eKit Components\u003c\/h2\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eComponent\u003c\/th\u003e\n\u003cth\u003eKit (enzyme not included), 100 assays (DSA100K)\u003c\/th\u003e\n\u003cth\u003eKit Plus (enzyme included), 100 assays (DSA100KE)\u003c\/th\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Buffer T2\u003c\/td\u003e\n\u003ctd\u003e600 µl\u003c\/td\u003e\n\u003ctd\u003e600 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x Relaxed DNA\u003c\/td\u003e\n\u003ctd\u003e405 µl\u003c\/td\u003e\n\u003ctd\u003e405 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003eH19 dye\u003c\/td\u003e\n\u003ctd\u003e8 µl (4500 x)\u003c\/td\u003e\n\u003ctd\u003e20 µl (1500 x)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x ATP\u003c\/td\u003e\n\u003ctd\u003e450 µl\u003c\/td\u003e\n\u003ctd\u003e450 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e10 x H19 dilution buffer\u003c\/td\u003e\n\u003ctd\u003e3000 µl\u003c\/td\u003e\n\u003ctd\u003e3000 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003ctd\u003e100 x E. coli gyrase\u003c\/td\u003e\n\u003ctd\u003eNot included\u003c\/td\u003e\n\u003ctd\u003e50 µl\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003cp\u003eFinal reaction conditions: 20 mM Tris-HCl pH 8, 35 mM NH4OAc, 4.6% glycerol, 1 mM DTT, 0.005% Brij35, 8 mM MgCl2, 25 µg\/ml relaxed plasmid DNA, 1 mM ATP and 20 nM gyrase. Magnesium is essential; EDTA should be avoided. A reaction without ATP can serve as negative control.\u003c\/p\u003e\n\u003ch2\u003eAssay Principle\u003c\/h2\u003e\n\u003cp\u003eRelaxed and supercoiled DNA give different fluorescence with the H19 dye: in the presence of magnesium, relaxed DNA suppresses the dye signal much more than supercoiled DNA. As gyrase converts relaxed plasmid into its supercoiled form, fluorescence (Ex 485 nm \/ Em 535 nm) rises; inhibitors keep the signal low.\u003c\/p\u003e\n\u003ch2\u003eSpecifications\u003c\/h2\u003e\n\u003ctable class=\"bhp-spec-table\"\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Target\u003c\/th\u003e\n\u003ctd\u003eE. coli gyrase DNA supercoiling\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eTarget Type\u003c\/th\u003e\n\u003ctd\u003eDNA Topoisomerase II (Gyrase)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eOrganism\u003c\/th\u003e\n\u003ctd\u003eE. coli\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Type\u003c\/th\u003e\n\u003ctd\u003eEnzyme Activity Assay; Inhibitor Screening Assay\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eDetection Method\u003c\/th\u003e\n\u003ctd\u003eFluorescence\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eDetection Wavelength\u003c\/th\u003e\n\u003ctd\u003eEx 485 nm \/ Em 535 nm\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003ePlate Format\u003c\/th\u003e\n\u003ctd\u003e96-well or 384-well plate\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eInstrument Required\u003c\/th\u003e\n\u003ctd\u003eFluorescence microplate reader\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eAssay Time\u003c\/th\u003e\n\u003ctd\u003eAbout 65 min (60 min reaction + 5 min dye incubation)\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eSize\u003c\/th\u003e\n\u003ctd\u003e100 assays\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eStorage\u003c\/th\u003e\n\u003ctd\u003ePlease contact us to confirm storage conditions.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003ctr\u003e\n\u003cth\u003eShipping\u003c\/th\u003e\n\u003ctd\u003eShipped frozen.\u003c\/td\u003e\n\u003c\/tr\u003e\n\u003c\/table\u003e\n\u003ch2\u003eProtocol\u003c\/h2\u003e\n\u003cp\u003eThe manufacturer's protocol describes the procedure and reaction volumes.\u003c\/p\u003e\n\u003cul\u003e\u003cli\u003eE. coli DNA Topoisomerase II (Gyrase) Assay (PDF, see Documents): Covers catalog # DSA100KE.\u003c\/li\u003e\u003c\/ul\u003e\n\u003ch2\u003eSafety \u0026amp; Handling\u003c\/h2\u003e\n\u003cdiv class=\"bhp-warning-box\"\u003eFor Research Use Only (RUO). Not for use in diagnostic or therapeutic procedures. The E. coli DNA Topoisomerase II (Gyrase) Assay Kit is shipped frozen. Follow standard laboratory safety practices when handling the reagents. Please contact us to confirm storage conditions.\u003c\/div\u003e\n\u003c\/div\u003e","brand":"ProFoldin","offers":[{"title":"Kit (enzyme not included)","offer_id":67794878333293,"sku":"DSA100K","price":825.0,"currency_code":"USD","in_stock":true},{"title":"Kit Plus (enzyme included)","offer_id":67794883051885,"sku":"DSA100KE","price":1047.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/gyrase2.jpg?v=1792791968","url":"https:\/\/www.ebiohippo.com\/products\/e-coli-dna-topoisomerase-ii-gyrase-assay-kit-bht16300067","provider":"BioHippo","version":"1.0","type":"link"}